目的 基于TLR4/ NF-κB/ NLRP3通路探讨左归降糖舒心方含药血浆对脂多糖（LPS）诱导小鼠J774A.1巨 噬细胞损伤及焦亡的作用及机制。方法 将分化完全的J774A.1巨噬细胞随机分为空白对照组（10%空白血浆）、 模型组（10 μg·mL-1 LPS +10%空白血浆）、5%中药含药血浆组（10 μg·mL-1 LPS+5%左归降糖舒心方含药血浆）、 10%中药含药血浆组（10 μg·mL-1LPS+10%左归降糖舒心方含药血浆）。除空白对照组外，其余各组以10 μg·mL-1 LPS处理，诱导巨噬细胞炎性损伤及细胞焦亡。按照分组条件干预后，采用ELISA法检测细胞上清中肿瘤坏死 因子α（TNF-α）、白细胞介素6（IL-6）、白细胞介素1β（IL-1β）和白细胞介素18（IL-18）含量；免疫荧光法检测 细胞中 NLRP3、ASC 蛋白共表达情况；Western Blot 法检测细胞 TLR4/NF-κB/NLRP3 通路及焦亡相关蛋白 的表达水平。结果 与空白对照组比较，模型组J774A.1细胞上清液中的TNF-α、IL-6、IL-1β、IL-18含量均 显著升高（P < 0.01）；细胞的 NLRP3、ASC 荧光强度显著增强，蛋白共表达明显上调；TLR4、p-NF-κB p65、 NLRP3、GSDMD-N、IL-1β蛋白表达显著上调（P < 0.01）。与模型组比较，5%、10%中药含药血浆组细胞上清 液中的 TNF-α、IL-6、IL-1β、IL-18 含量均显著降低（P < 0.01）；细胞的 NLRP3、ASC 荧光强度均明显减弱， 蛋白共表达明显下调；TLR4、p-NF-κB p65、NLRP3、GSDMD-N、IL-1β 蛋白表达显著下调（P < 0.01）。 结论 左归降糖舒心方含药血浆能够抑制LPS诱导的J774A.1巨噬细胞损伤及炎症反应，其作用机制可能与调 控TLR4/NF-κB/NLRP3炎症通路，抑制细胞焦亡及炎症因子表达有关。

Objective To investigate the effect and mechanism of Zuogui Jiangtang Shuxin Formula-containing plasma on LPS-induced macrophage damage and pyroptosis based on Toll-like receptor 4（TLR4）/nuclear factor-κB（NF-κB）/ NOD-like receptor thermal protein domain associated protein 3 （NLRP3） pathway. Methods Fully differentiated J774A.1 cells were randomly divided into the blank control group（10% blank plasma），the model group（10 μg·mL-1 LPS +10% blank plasma），5%（10 μg·mL-1 LPS + 5% Zuogui Jiangtang Shuxin Formula-containing plasma）and 10% （10 μg · mL-1 LPS + 10% Zuogui Jiangtang Shuxin Formula-containing plasma） Zuogui Jiangtang Shuxin Formula-containing plasma group. Except for the blank control group，other groups were treated with 10 μg·mL-1 LPS to induce inflammatory injury and pyroptosis of macrophage. After the intervention， ELISA was used to detect concentration of tumor necrosis factor-alpha（TNF - α）， interleukin 6（IL-6）， interleukin 1 beta（IL-1β）and interleukin 18（IL-18）in macrophage supernatant. The co-expression of NOD-like receptor protein 3（NLRP3）and apoptotic spot-like protein（ASC）were detected by immunofluorescence assay. The expression of TLR4/NF-κB/NLRP3 signaling pathway and pyroptosis-related proteins were detected by Western Blot. Results Compared with the blank control group， the levels of TNF - α， IL-6， IL-1β and IL-18 in the supernatant of LPS-treated J774A. 1 cells significantly increased（P < 0.01）. It was also found that the fluorescence intensity of NLRP3 and ASC in cells was significantly enhanced，and protein co-expression was significantly up-regulated. The protein co-expressions of TLR4， p-NF-κB p65，NLRP3，GSDMD-N and IL-1β were significantly up-regulated（P < 0.01）. Compared with the model group， the levels of TNF - α， IL-6， IL-1β and IL-18 in the supernatant of Zuogui Jiangtang Shuxin Formulacontaining plasma-treated J774A.1 cells were obviously decreased（P < 0.01）. The fluorescence intensity of NLRP3 and ASC in cells was significantly reduced，and protein co-expression was significantly down-regulated. The protein coexpressions of TLR4，p-NF-κB p65，NLRP3，GSDMD-N and IL-1β were significantly down-regulated（P < 0.01）. Conclusion Zuogui Jiangtang Shuxin Formula-containing plasma can suppresses LPS-induced macrophage damage and inflammatory response，and its mechanism may be related to the regulation of TLR4 / NF - κB / NLRP3 signaling pathway and inhibition of cellular pyroptosis and inflammatory factor expression.

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国家自然科学基金项目（82205077）；湖南省自然科学基金项目（2022JJ40315）；湖南省教育厅资助科研项目（21B0390）；湖南省卫生 健康委科研计划项目（D202303067601）；中药粉体与创新药物省部共建国家重点实验室培育基地开放基金项目（21PTKF1016）